Heterocyclic inhibitors of lysine biosynthesis via the diaminopimelic acid pathway
阅读说明：本技术 通过二氨基庚二酸途径的赖氨酸生物合成的杂环抑制剂 (Heterocyclic inhibitors of lysine biosynthesis via the diaminopimelic acid pathway ) 是由 马修·A·佩鲁吉尼 别林达·阿沃特 塔蒂亚娜·苏亚雷斯达科斯塔 于 2018-04-12 设计创作，主要内容包括：本发明涉及式(1)的某些杂环化合物,其能够在某些生物中抑制通过二氨基庚二酸生物合成途径的赖氨酸生物合成。作为这种活性的结果,这些化合物可用于其中抑制赖氨酸生物合成有用的应用。这种类型的应用包括将所述化合物用作除草剂。<Image he="482" wi="606" file="DDA0002269436250000011.GIF" imgContent="drawing" imgFormat="GIF" orientation="portrait" inline="no"></Image>(The present invention relates to certain heterocyclic compounds of formula (1) which are capable of inhibiting lysine biosynthesis via the diaminopimelic acid biosynthetic pathway in certain organisms. As a result of this activity, these compounds are useful in applications where inhibition of lysine biosynthesis is useful. This type of application involves the use of the compounds as herbicides.)
1. A method of inhibiting lysine biosynthesis in an organism in which the diaminopimelic acid biosynthesis pathway occurs, said method comprising contacting said organism with an effective amount of a compound of formula (I):
wherein:
X、X1and X2Each independently selected from O, NH and S;
ar is optionally substituted C6-C18Aryl or optionally substituted C1-C18A heteroaryl group;
each R is H, or two R when taken together form a double bond between the carbon atoms to which they are attached;
l is selected from the group consisting of a bond, C1-C6Alkyl radical, C2-C6Alkenyl radical, C1-C6Alkoxy radical, C1-C6Alkoxy radical C1-C6Alkyl and C1-C6A heteroalkyl group;
R1selected from H, OH, CN, tetrazole, CO2H and COR2；
R2Selected from H, Cl, NR3R4、O-C1-C6Alkyl and O-C1-C6A heteroalkyl group;
R3and R4Each independently selected from H and C1-C6An alkyl group.
2. The method of claim 1, wherein in the compound of formula I, two R when taken together form a double bond between the carbon atoms to which they are attached.
3. The method of claim 1 or 2, wherein in the compound of formula I, X is S.
4. A method according to any one of claims 1 to 3, wherein in the compound of formula I, X1Is O.
5. The method according to any one of claims 1 to 4, wherein in the compound of formula I, wherein X is2Is O.
6. The method according to any one of claims 1 to 5, wherein in the compound of formula I, Ar is selected from:
A1、A2、A3、A4and A5Each independently selected from N and CR5；
V1、V2、V3And V4Each independently selected from N and CR5；
Y is selected from S, O and NH;
each R5Independently selected from H, halogen, OH, NO2、CN、SH、NH2、CF3、OCF3、C1-C12Alkyl radical, C1-C12Alkoxy radical, C1-C12Haloalkyl, C2-C12Alkenyl radical, C2-C12Alkynyl, C2-C12Heteroalkyl, SR6、SO3H、SO2NR6R6、SO2R6、SONR6R6、SOR6、COR6、COOH、COOR6、CONR6R6、NR6COR6、NR6COOR6、NR6SO2R6、NR6CONR6R6、NR6R6And an acyl group;
or any two R on adjacent carbon atoms5When taken together with the carbon atom to which they are attached form a 5 or 6 membered ring moiety;
each R6Independently selected from H and C1-C12An alkyl group.
7. The method of any one of claims 1 to 6, wherein in the compound of formula I, Ar has the formula:
wherein A is1、A2、A3、A4And A5As defined in claim 6.
8. The method according to any one of claims 1 to 7, wherein in the compound of formula I, Ar is selected from:
9. the method of any one of claims 6 to 8, wherein in the compound of formula I, each R is5Independently selected from H, Cl, Br, F, OH, NO2、NH2、C1-C12Alkyl radical, C1-C12Alkoxy and NR6COR6。
10. The method of any one of claims 6 to 9, wherein in the compound of formula I, each R is5Independently selected from H, F, Cl, Br, I, CH3、CH2CH3、CH2NH2、OH、OCH3、SH、SCH3、CO2H、CONH2、CF3、OCF3、NO2、NH2CN and NHCOCH.
11. The method according to any one of claims 1 to 10, wherein in the compound of formula I, L is C1-C6An alkyl group.
12. The method of claim 11, L is C of the formula1-C6Alkyl groups:
-(CH2)a-；
wherein a is selected from 1, 2, 3 and 4.
13. The method of claim 12, wherein in the compound of formula I, a is 1.
14. Any of claims 1 to 13A method of the formula I, wherein in the compound of the formula I, R1Is CO2H。
15. The method of any one of claims 1 to 14, wherein the organism is a plant.
16. The method of any one of claims 1 to 15, wherein the compound inhibits lysine biosynthesis in the organism by inhibiting the Diaminopimelic Acid (DAP) pathway.
17. The method of any one of claims 1 to 16, wherein the compound inhibits lysine biosynthesis in the organism by inhibiting DHDPS activity.
18. The method of any one of claims 1 to 17, wherein the compound is selected from the group consisting of:
19. a method for controlling undesired plant growth, which comprises contacting the plants with a herbicidally effective amount of a compound of formula (I):
wherein:
X、X1and X2Each independently selected from O, NH and S;
ar is optionally substituted C6-C18Aryl or optionally substituted C1-C18A heteroaryl group;
each R is H, or two R when taken together form a double bond between the carbon atoms to which they are attached;
l is selected from the group consisting of a bond, C1-C6Alkyl radical, C2-C6Alkenyl radical, C1-C6Alkoxy radical, C1-C6Alkoxy radical C1-C6Alkyl and C1-C6A heteroalkyl group;
R1selected from H, OH, CN, tetrazole, CO2H and COR2；
R2Selected from H, Cl, NR3R4、O-C1-C6Alkyl and O-C1-C6A heteroalkyl group;
R3and R4Each independently selected from H and C1-C6An alkyl group.
20. The method of claim 19, wherein in the compound of formula I used in the method, two R when taken together form a double bond between the carbon atoms to which they are attached.
21. The method of claim 19 or 20, wherein in the compound of formula I used in the method, X is S.
22. A method according to any one of claims 19 to 21, wherein in the compound of formula I used in the method, X1Is O.
23. A method according to any one of claims 19 to 22, wherein in the compound of formula I used in the method, X2Is O.
24. A process according to any one of claims 19 to 23, wherein in the compound of formula I used in the process, Ar is selected from:
A1、A2、A3、A4and A5Each independently selected from N and CR5；
V1、V2、V3And V4Each independently selected from N and CR5；
Y is selected from S, O and NH;
each R5Independently selected from H, halogen, OH, NO2、CN、SH、NH2、CF3、OCF3、C1-C12Alkyl radical, C1-C12Alkoxy radical, C1-C12Haloalkyl, C2-C12Alkenyl radical, C2-C12Alkynyl, C2-C12Heteroalkyl, SR6、SO3H、SO2NR6R6、SO2R6、SONR6R6、SOR6、COR6、COOH、COOR6、CONR6R6、NR6COR6、NR6COOR6、NR6SO2R6、NR6CONR6R6、NR6R6And an acyl group;
or any two R on adjacent carbon atoms5When taken together with the carbon atom to which they are attached form a 5 or 6 membered ring moiety;
each R6Independently selected from H and C1-C12An alkyl group.
25. A process according to any one of claims 19 to 24, wherein in the compound of formula I used in the process, Ar has the formula:
wherein A is1、A2、A3、A4And A5As defined in claim 28.
26. A process according to any one of claims 19 to 25, wherein in the compound of formula I used in the process, Ar is selected from:
27. the method of any one of claims 24 to 26, wherein in the compound of formula I used in the method, each R is5Independently selected from H, Cl, Br, F, OH, NO2、NH2、C1-C12Alkyl radical, C1-C12Alkoxy and NR6COR6。
28. The method of any one of claims 24 to 27, wherein in the compound of formula I used in the method, each R is5Independently selected from H, F, Cl, Br, I, CH3、CH2CH3、CH2NH2、OH、OCH3、SH、SCH3、CO2H、CONH2、CF3、OCF3、NO2、NH2CN and NHCOCH.
29. A method according to any one of claims 19 to 28, wherein in the compound of formula I used in the method, L is C1-C6An alkyl group.
30. A process according to claim 29, wherein in the compound of formula I used in the process, L is C of formula1-C6Alkyl groups:
-(CH2)a-；
wherein a is selected from 1, 2, 3 and 4.
31. The method of claim 30, wherein in the compound of formula I used in the method, a is 1.
32. The method of claim 30 or 31, wherein in the compound of formula I used in the method, R is1Is CO2H。
33. The method according to any one of claims 19 to 28, wherein the compound used in the method is selected from the group consisting of:
34. the method of any one of claims 19 to 33, wherein the plant is contacted with the compound by spraying the plant with a composition comprising the compound.
35. The method of claim 34, wherein the composition comprises from 1 Wt% to 90 Wt% of the compound of formula 1.
Technical Field
The present invention relates to substituted heterocyclic compounds capable of inhibiting lysine biosynthesis via the diaminopimelate pathway in certain organisms. As a result of this activity, these compounds are useful in applications where inhibition of lysine biosynthesis is useful. This type of application includes the use of the compounds as herbicides.
Background
Chemical agents have been widely used in many applications over the 20 th century, including as pharmaceuticals, herbicides, pesticides, and the like. Unfortunately, due to the widespread use of these agents, many compounds that exhibit useful activity no longer function because the target species develops some form of resistance to the active agent.
The development and use of herbicides has a significant impact on the ability to support an ever-increasing world population. Herbicides have helped farmers perform weed management of crops and have also promoted no-tillage crop production to preserve soil and moisture. Thus, its use has had a significant positive impact on crop yield and productivity per hectare.
Unfortunately, repeated application of herbicides with the same mechanism of action to crops or fields has resulted in the production of herbicide resistant weeds. It is believed that weeds are herbicide resistant due to herbicide selection pressure by which, once the herbicide has been applied, those weeds that have some form of resistance are favored, resulting in a selective advantage over resistant weeds.
Recognizing the development of herbicide resistance, there is a continuing need to develop new agents that can be used as alternative active agents to those that no longer function in the field due to resistance development. Accordingly, there is a continuing need to develop new compounds that can be used as herbicides or to identify existing compounds that can be used as herbicides.
One challenge in developing an active agent that is a herbicide is to ensure that the developed agent has an acceptable safety profile after exposure to humans, since ideally the agent would be non-toxic or extremely low-toxic to humans (and preferably mammals) as a whole.
In this regard, an attractive target for the development of this type of reagent is the biosynthesis of the amino acid lysine and its direct precursor meso-diaminopimelate (meso-DAP). This is an attractive route of investigation, since the lysine biosynthetic pathway occurs in plants and bacteria, but not in mammals. Mammals lack the ability to biosynthetically produce lysine, and thus lysine is one of the 9 essential amino acids that must be provided from dietary sources. The occurrence of the lysine biosynthetic pathway in plants and the absence of specific inhibitors in mammals indicates that specific inhibitors of this biosynthetic pathway will exhibit novel activity and low mammalian toxicity.
Therefore, it would be desirable to develop inhibitors of the lysine biosynthetic pathway, as it is expected that these inhibitors would potentially have interesting herbicidal activity.
Disclosure of Invention
Accordingly, the applicant of the present invention has conducted studies on the diaminopimelic acid pathway to identify inhibitors of lysine biosynthesis that could potentially be applied as herbicides.
As a result of these studies, applicants have identified compounds having the ability to inhibit lysine biosynthesis.
Accordingly, in one embodiment, the present invention provides a method of inhibiting lysine biosynthesis in an organism in which the diaminopimelic acid biosynthesis pathway occurs, the method comprising contacting the organism with an effective amount of a compound of formula (1):
wherein:
X、X1and X2Each independently selected from O, NH and S;
ar is optionally substituted C6-C18Aryl or optionally substituted C1-C18A heteroaryl group;
each R is H, or two R when taken together form a double bond between the carbon atoms to which they are attached;
l is selected from the group consisting of a bond, C1-C6Alkyl radical, C2-C6Alkenyl radical, C1-C6Alkoxy radical, C1-C6Alkoxy radical C1-C6Alkyl and C1-C6A heteroalkyl group;
R1selected from H, OH, CN, tetrazole, CO2H and COR2；
R2Selected from H, Cl, NR3R4、O-C1-C6Alkyl and O-C1-C6A heteroalkyl group;
R3and R4Each independently selected from H and C1-C6An alkyl group.
Without wishing to be bound by theory, it is believed that the compounds have activity in inhibiting lysine biosynthesis in organisms by inhibiting the Diaminopimelic Acid (DAP) pathway. In particular, the compounds are believed to inhibit the dihydrodipicolinate synthase (DHDPS) pathway in an organism by inhibiting this pathway.
Since the compounds are capable of inhibiting the lysine biosynthetic pathway, applicants have also discovered that the compounds are useful as herbicides, since the lysine biosynthetic pathway is an essential pathway in plants.
Thus, in another aspect, the present invention provides a method for controlling (control) undesired plant growth, which comprises contacting the plants with a herbicidally effective amount of a compound of formula (1):
wherein:
X、X1and X2Each independently selected from O, NH and S;
ar is optionally substituted C6-C18Aryl or optionally substituted C1-C18A heteroaryl group;
each R is H, or two R when taken together form a double bond between the carbon atoms to which they are attached;
l is selected from the group consisting of a bond, C1-C6Alkyl radical, C2-C6Alkenyl radical, C1-C6Alkoxy radical, C1-C6Alkoxy radical C1-C6Alkyl and C1-C6A heteroalkyl group;
R1selected from H, OH, CN, tetrazole, CO2H and COR2；
R2Selected from H, Cl, NR3R4、O-C1-C6Alkyl and O-C1-C6A heteroalkyl group;
R3and R4Each independently selected from H and C1-C6An alkyl group.
Brief Description of Drawings
FIG. 1 shows the diaminopimelic acid biosynthesis pathway in bacteria and plants.
Fig. 2 shows the structures of meso-dap (a) and lysine (B).
FIG. 3 shows the first step in the diaminopimelic acid biosynthetic pathway catalyzed by DHDPS.
Figure 4 shows the following DHDPS enzyme structures: the head-to-head dimer observed for most bacterial species (a), the back-to-back dimer observed for plant species (B), and the dimer form observed for certain bacterial species (C), where a, B, C, and d refer to the monomeric units of the protein.
Figure 5 shows a plot of root length versus concentration for plants treated with (a) compound 3 and (b) compound 5.
Detailed Description
In this specification, a number of terms are used which are well known to the skilled person. However, for the sake of clarity, a number of terms will be defined.
Throughout the description of this specification and the claims that follow, the word "comprise" and variations thereof are not intended to exclude other additional items, components, integers or steps.
The term "effective amount" means an amount sufficient to achieve the desired beneficial result. For herbicides, an effective amount is an amount sufficient to control undesired plant growth.
The term "inhibit" and variations thereof mean to prevent, block or reduce the function of an inhibited object. The term does not require complete inhibition, wherein an activity reduction of at least 50% is considered inhibition.
The term "control" in relation to plant growth means reducing or eliminating the growth of a plant. This may involve killing the plant but also includes within its scope hindering or reducing plant growth.
The term "or a salt thereof" refers to a salt that retains the desired biological activity of the identified compound described above and includes acid addition salts and base addition salts. Suitable acceptable acid addition salts of the compounds of formula (1) may be prepared from inorganic acids or from organic acids. Some examples of such mineral acids are hydrochloric acid, sulfuric acid and phosphoric acid. Suitable organic acids may be selected from: aliphatic, alicyclic, aromatic, heterocyclic, carboxylic and sulfonic organic acids, some examples of which are formic, acetic, propionic, pyruvic, succinic, glycolic, gluconic, lactic, malic, tartaric, citric, fumaric, maleic, alkylsulfonic and arylsulfonic acids. Additional information on pharmaceutically acceptable Salts can be found in P.H.Stahl and C.G.Wermuth Handbook of Pharmaceutical Salts, Properties, Selection, and Use, revision 2, Wiley-VCH 2011. In the case of reagents that are solids, those skilled in the art will appreciate that the compounds, reagents and salts may exist in different crystalline or polymorphic forms, all of which are intended to be within the scope of the invention and the specified formula.
The term "optionally substituted" as used throughout the specification means that a group may or may not be further substituted with one or more non-hydrogen substituents or fused thereto (to form a condensed polycyclic system). In certain embodiments, the substituents are one or more groups independently selected from: halogen, ═ O, ═ S, -CN, -NO2、-CF3、-OCF3Alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl, cycloalkylalkyl, heterocycloalkylalkyl, heteroarylalkyl, arylalkyl, cycloalkylalkenyl, heterocycloalkylalkenyl, arylalkenyl, heteroarylalkenyl, cycloalkylheteroalkyl, heterocycloalkylheteroalkyl, arylheteroalkyl, heteroarylheteroalkyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, alkoxycycloalkyl, alkoxyheterocycloalkyl, alkoxyaryl, alkoxyheteroaryl, alkoxycarbonyl, alkylaminocarbonyl, alkenyloxy, alkynyloxy, cycloalkoxy, cycloalkenyloxy, heterocycloalkoxy, heterocycloalkenyloxy, aryloxyaloxy, aryloxyalkyl, alkoxycycloalkyl, alkoxyalkoxy-heteroaryl, alkoxyalkoxy-carbonyl, alkylaminocarbonyl, alkenyloxy, alkynyloxy, cycloalkenyloxy, heterocycloalkyloxy, heterocycloalkenyloxy, and cycloalkenyloxyA group, phenoxy, benzyloxy, heteroaryloxy, arylalkoxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, sulfinyl, alkylsulfinyl, arylsulfinyl, aminosulfinylaminoalkyl, C (═ O) OH, -C (═ O) Re、C(＝O)ORe、C(＝O)NReRf、C(＝NOH)Re、C(＝NRe)NRfRg、NReRf、NReC(＝O)Rf、NReC(＝O)ORf、NReC(＝O)NRfRg、NReC(＝NRf)NRgRh、NReSO2Rf、-SRe、SO2NReRf、-ORe、OC(＝O)NReRf、OC(＝O)ReAnd an acyl group, and a salt thereof,
wherein R ise、Rf、RgAnd RhEach independently selected from: H. c1-C12Alkyl radical, C1-C12Haloalkyl, C2-C12Alkenyl radical, C2-C12Alkynyl, C1-C10Heteroalkyl group, C3-C12Cycloalkyl radical, C3-C12Cycloalkenyl radical, C1-C12Heterocyclylalkyl radical, C1-C12Heterocycloalkenyl, C6-C18Aryl radical, C1-C18Heteroaryl and acyl; or Re、Rf、RgAnd RhAny two or more of which, when taken together with the atoms to which they are attached, form a heterocyclic ring system having from 3 to 12 ring atoms.
Some examples of particularly suitable optional substituents include F, Cl, Br, I, CH3、CH2CH3、CH2NH2、OH、OCH3、SH、SCH3、CO2H、CONH2、CF3、OCF3、NO2、NH2And CN.
In the following definitions of the various substituents, it is stated that "a group may be a terminal group or a bridging group". This is intended to mean that the use of this term is intended to encompass the case where the group is a linker between two other parts of the molecule and where it is a terminal moiety. Using the term alkyl as an example, some publications use the term "alkylene" for the bridging group, and thus in these other publications there is a distinction between the term "alkyl" (end group) and "alkylene" (bridging group). In the present application, no such distinction is made, and most groups may be bridging or terminal groups.
"alkenyl" as a group or part of a group denotes an aliphatic hydrocarbon group containing at least one carbon-carbon double bond, and which may be straight or branched, preferably having from 2 to 12 carbon atoms, more preferably from 2 to 10 carbon atoms, most preferably from 2 to 6 carbon atoms in the normal chain. The group may contain multiple double bonds in the normal chain and is independently E or Z with respect to the orientation of each. Alkenyl is preferably 1-alkenyl. Exemplary alkenyl groups include, but are not limited to, ethenyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, and nonenyl. The groups may be terminal groups or bridging groups.
Unless otherwise indicated, "alkyl" as a group or part of a group refers to a straight or branched chain aliphatic hydrocarbon group, preferably C1-C12Alkyl, more preferably C1-C10Alkyl, most preferably C1-C6. Suitable straight and branched chains C1-C6Some examples of alkyl substituents include methyl, ethyl, n-propyl, 2-propyl, n-butyl, sec-butyl, tert-butyl, hexyl and the like. The groups may be terminal groups or bridging groups.
"alkoxy" refers to an alkyl-O-group, wherein alkyl is as defined herein. Preferably, alkoxy is C1-C6An alkoxy group. Examples include, but are not limited to, methoxy and ethoxy. The groups may be terminal groups or bridging groups.
"alkoxyalkyl" refers to an alkoxy-alkyl-group, wherein the alkoxy and alkyl portions are as defined herein. The groups may be terminal groups or bridging groups. If the group is a terminal group, it is bonded to the rest of the molecule through an alkyl group.
"aryl" as a group or part of a group means (i) an optionally substituted monocyclic or fused polycyclic aromatic carbocyclic ring (ring structures in which the ring atoms are all carbon), preferably having 5 to 12 atoms per ring. Some examples of aryl groups include phenyl, naphthyl, and the like; (ii) an optionally substituted partially saturated bicyclic aromatic carbocyclic moiety wherein phenyl and C5-7Cycloalkyl or C5-7Cycloalkenyl groups are fused together to form a ring structure, such as tetrahydronaphthyl, indenyl, or indanyl. The groups may be terminal groups or bridging groups. In general, the aryl group is C6-C18And (4) an aryl group.
"heteroalkyl" means a straight or branched chain alkyl group preferably having 2 to 12 carbons, more preferably 2 to 6 carbons in the chain, in which one or more carbon atoms (and any associated hydrogen atoms) are each independently replaced with a heteroatom group selected from S, O, P and NR ', wherein R' is selected from H, optionally substituted C1-C12Alkyl, optionally substituted C3-C12Cycloalkyl, optionally substituted C6-C18Aryl and optionally substituted C1-C18A heteroaryl group. Exemplary heteroalkyl groups include alkyl ethers, secondary and tertiary alkyl amines, amides, alkyl sulfides, and the like. Some examples of heteroalkyl groups also include hydroxy C1-C6Alkyl radical, C1-C6Alkoxy radical C1-C6Alkyl, amino C1-C6Alkyl radical, C1-C6Alkylamino radical C1-C6Alkyl and di (C)1-C6Alkyl) amino C1-C6An alkyl group. The groups may be terminal groups or bridging groups.
"heteroaryl" alone or as part of a group refers to a group comprising an aromatic ring (preferably a 5 or 6 membered aromatic ring) having one or more heteroatoms as ring atoms in the aromatic ring, the remaining ring atoms being carbon atoms. Suitable heteroatoms include nitrogen, oxygen, and sulfur. Some examples of heteroaryl groups include: thiophene; benzothiophenes; a benzofuran; benzimidazole; benzo (b) isAzole; benzothiazole; benzisothiazole; naphtho [2, 3-b ]]Thiophene; furan; isoindoline; xanthone (xantholene); phenOxazines (phenoxatine); pyrrole; imidazole; pyrazole; pyridine; pyrazine; a pyrimidine; pyridazine; tetrazole; indole; isoindole; 1H-indazole; a purine; quinoline; isoquinoline; phthalazine; naphthyridine; quinoxaline; cinnoline; carbazole; phenanthridine; acridine; a phenazine; a thiazole; isothiazole; phenothiazine;azole; different from each otherAzole; furazan (furazane); phenAn oxazine; 2-, 3-or 4-pyridyl; 2-, 3-, 4-, 5-, or 8-quinolinyl; 1-, 3-, 4-or 5-isoquinolinyl; 1-, 2-or 3-indolyl, and 2-or 3-thienyl. Heteroaryl is usually C1-C18A heteroaryl group. The groups may be terminal groups or bridging groups.
As shown in fig. 1, lysine synthesis in bacteria via the diaminopimelic acid pathway begins with the combination of Pyruvate (PYR) and L-Aspartate Semialdehyde (ASA) in the presence of dihydrodipicolinate synthase (DHDPS) to synthesize 2, 3, 4, 5-tetrahydro-L, L-dipicolinic acid (HTPA). The HTPA will be dehydrated and the dihydropyridine dicarboxylic acid (DHDP) will be generated by a non-enzymatic step. DHDP will be reduced by the enzyme dihydrodipicolinate reductase (DHDPR), which is an nad (p) H-dependent enzyme, to form 2, 3, 4, 5-tetrahydropyridinedicarboxylic acid (2, 3, 4, 5-tetrahydrodipicolinate, THDP). Then, THDP will go through one of four pathways; succinylase (succinylase), acetylase (acetylase), dehydrogenase or aminotransferase, depending on the species of bacteria and plants. All routes lead to the synthesis of the common, biologically important compound meso-L, L' -2, 6-diaminopimelic acid (meso-DAP). meso-DAP is subsequently decarboxylated by the enzyme diaminopimelate decarboxylase (DAPDC), leading to the formation of lysine. The generated meso-DAP serves as a cross-linking moiety in the peptidoglycan layer of the cell wall of gram-negative as well as gram-positive bacteria, such as Bacillus (Bacillus sp). Lysine also forms peptidoglycan cross-links in the bacterial cell wall of most gram-positive bacteria and is used for protein synthesis in both bacteria and plants. Thus, lysine is essential for both bacterial and plant cell function and viability.
Referring to FIG. 1, the first step of the diaminopimelate biosynthetic pathway requires the enzyme dihydrodipicolinate synthase (DHDPS). An expanded view of this first step is shown in fig. 3. As can be seen, this step involves combining Pyruvate (PYR) and L-Aspartate Semialdehyde (ASA) in the presence of dihydrodipicolinate synthase (DHDPS) to form 2, 3, 4, 5-tetrahydro-L, L-dipicolinic acid (HTPA). Since this step in the diaminopimelic acid biosynthetic pathway is common to all bacteria and plants, it is believed to provide an attractive target in the development of inhibitors of lysine biosynthesis.
The enzyme dihydrodipicolinate synthase (DHDPS) was characterized in 1965 after purification from Escherichia coli (e.coli). After the enzyme has been characterized, extensive research has been conducted on it, in which crystal structure work of the enzyme has been conducted.
As can be seen from fig. 4, the quaternary structure of DHDPS in gram-negative bacteria consists of four monomer units linked together in such a way that only one monomer interacts with two other monomers (fig. 4A). The tetrameric structure, which is also referred to as a "head-to-head" dimer-dimer, has large cavities filled with water. As shown in fig. 4A, two monomers interact more tightly than the other two monomers, and thus are referred to as a tight dimer interface and a weak dimer interface, respectively. The active site of the enzyme is located at the tight dimer interface. In the active site of E.coli, threonine 44 and tyrosine 133 are present, with tyrosine 107 crossing over the two monomers at the interface of the tight dimers, thus allowing two active sites to be generated per dimer.
The structure of DHDPS in plants also consisted of tetramers, but the structure appeared to be "back-to-back" dimer-dimer (fig. 4B). DHDPS in certain bacterial species, such as Staphylococcus aureus (Staphylococcus aureus) and Pseudomonas aeruginosa (Pseudomonas aeruginosa), exists only as dimers consisting of tightly bound dimer interfaces (fig. 4C).
It can be seen that the first step in the diaminopimelic acid biosynthetic pathway is common in plants and therefore represents an attractive target for compound development in the field of herbicides.
As discussed above, the applicant of the present invention has identified compounds capable of inhibiting lysine biosynthesis via the diaminopimelic pathway. Accordingly, in one embodiment, the present invention provides a method of inhibiting lysine biosynthesis in an organism in which the diaminopimelic acid biosynthesis pathway occurs, said method comprising contacting the organism with an effective amount of a compound of formula (I). The person skilled in the art will readily understand the organisms in which the diaminopimelic acid biosynthetic pathway takes place. However, for the avoidance of doubt, we point out that all species in the kingdoms Archaea (Archaea), Eubacteria (both gram-negative and gram-positive species) and plants (from moss species up to higher plants) utilise the diaminopimelic acid pathway and are therefore considered to be organisms in which the diaminopimelic acid pathway occurs.
The compounds used in the process of the invention are compounds of formula (1) or salts or N-oxides thereof:
wherein:
X、X1and X2Each independently selected from O, NH and S;
ar is optionally substituted C6-C18Aryl radicalsOr optionally substituted C1-C18A heteroaryl group;
each R is H, or two R when taken together form a double bond between the carbon atoms to which they are attached;
l is selected from the group consisting of a bond, C1-C6Alkyl radical, C2-C6Alkenyl radical, C1-C6Alkoxy radical, C1-C6Alkoxy radical C1-C6Alkyl and C1-C6A heteroalkyl group;
R1selected from H, OH, CN, tetrazole, CO2H and COR2；
R2Selected from H, Cl, NR3R4、O-C1-C6Alkyl and O-C1-C6A heteroalkyl group;
R4and R5Each independently selected from H and C1-C6An alkyl group.
In the compounds used in the method of the present invention, each R is H; or two R when taken together form a double bond between the carbon atoms to which they are attached. In one embodiment, each R is H. In one embodiment, two R groups, when taken together, form a double bond between the carbon atoms to which they are attached. This provides the compound of formula (2).
Wherein Ar, X1、X2L and R1As defined above.
Theoretically, the geometry around the double bond in the compound of formula (2) can be E or Z. In one embodiment, the compound is the E isomer. In one embodiment, the geometric structure is the Z isomer. In one embodiment, the geometry is such that the compound is a compound of formula (3):
wherein Ar, X1、X2L andR1as defined above.
Of the compounds used in the process of the present invention, X, X1And X2Each independently selected from O, NH and S.
In one embodiment, X is S. In one embodiment, X is O. In one embodiment, X is NH. In one embodiment, X1Is S. In one embodiment, X1Is O. In one embodiment, X1Is NH. In one embodiment, X2Is S. In one embodiment, X2Is O. In one embodiment, X2Is NH. As one skilled in the art will appreciate, since there are three potential values for each variable, there are 27 possible combinations, all of which are intended to be covered by this application.
In one embodiment of the compounds of formula (3) used in the methods of the present invention, X is S, providing a compound of formula (3 a):
wherein Ar and X1、X2L and R1As defined above.
In one embodiment of the compounds of formula (3) used in the methods of the present invention, X is O, providing a compound of formula (3 b):
wherein Ar and X1、X2L and R1As defined above.
In one embodiment of the compounds of formula (3) used in the methods of the present invention, X is NH, providing a compound of formula (3 c):
wherein Ar and X1、X2L and R1As defined above.
In one embodiment of the compounds of formula (3a) used in the process of the invention, X1To O, there is provided a compound of formula (3 aa):
wherein Ar and X2L and R1As defined above.
In one embodiment of the compounds of formula (3b) used in the process of the present invention, X1To O, a compound of formula (3 ba):
wherein Ar and X2L and R1As defined above.
In one embodiment of the compounds of formula (3c) used in the process of the invention, X1To O, there is provided a compound of formula (3 ca):
wherein Ar and X2L and R1As defined above.
In one embodiment of the compound of formula (3aa) used in the process of the present invention, X2To O, there is provided a compound of formula (3 aaa):
wherein Ar, L and R1As defined above.
In one embodiment of the compound of formula (3ba) used in the method of the present invention, X2To O, compounds of formula (3baa) are provided:
wherein Ar, L andR1as defined above.
In one embodiment of the compound of formula (3ca) used in the process of the invention, X2To O, there is provided a compound of formula (3 caa):
wherein Ar, L and R1As defined above.
In the compounds used in the process of the invention, Ar is optionally substituted C6-C18Aryl or optionally substituted C1-C18A heteroaryl group.
In some embodiments, the group Ar is optionally substituted C6-C18And (4) an aryl group. Some examples of such groups include optionally substituted phenyl and optionally substituted naphthyl.
In some embodiments, the group Ar may be any optionally substituted C1-C18A heteroaryl group. Suitable heteroaryl groups include: thiophene; benzothiophenes; a benzofuran; benzimidazole; benzo (b) isAzole; benzothiazole; benzisothiazole; naphtho [2, 3-b ]]Thiophene; furan; isoindoline; xanthone; phenAn oxazine; pyrrole; imidazole; pyrazole; pyridine; pyrazine; a pyrimidine; pyridazine; tetrazole; indole; isoindole; 1H-indazole; a purine; quinoline; isoquinoline; phthalazine; naphthyridine; quinoxaline; cinnoline; carbazole; phenanthridine; acridine; a phenazine; a thiazole; isothiazole; phenothiazine;azole; different from each otherAzole; furazan; phenAn oxazine; a pyridyl group; a quinolyl group; an isoquinolinyl group; indolyl and thienyl. In each case where multiple substitution sites are possible on the heteroaryl ring, all possible points of attachment are contemplated. By way of example only, if heteroaryl is a pyridyl moiety, it may be 2-pyridyl, 3-pyridyl, or 4-pyridyl.
In some embodiments, Ar is selected from:
wherein A is1、A2、A3、A4And A5Each independently selected from N and CR5；
V1、V2、V3And V4Each independently selected from N and CR5；
Y is selected from S, O and NH;
each R5Independently selected from H, halogen, OH, NO2、CN、SH、NH2、CF3、OCF3、C1-C12Alkyl radical, C1-C12Alkoxy radical, C1-C12Haloalkyl, C2-C12Alkenyl radical, C2-C12Alkynyl, C2-C12Heteroalkyl group, C6-C18Aryl radical C1-C12Alkoxy, SR6、SO3H、SO2NR6R6、SO2R6、SONR6R6、SOR6、COR6、COOH、COOR6、CONR6R6、NR6COR6、NR6COOR6、NR6SO2R6、NR6CONR6R6、NR6R6And an acyl group;
or any two R on adjacent carbon atoms5When taken together with the carbon atom to which they are attached form a 5 or 6 membered ring moiety;
each R6Independently selected from H and C1-C12An alkyl group.
In some embodiments, Ar is an aromatic moiety of the formula:
wherein A is1、A2、A3、A4And A5As defined above.
In some embodiments, Ar is an aromatic moiety selected from the group consisting of:
in some embodiments, Ar is selected from:
wherein V1、V2、V3And V4Each independently selected from N and CR5；
Y is selected from S, O and NH.
In one embodiment, Ar is selected from:
wherein R is5As described above.
In one embodiment, Ar is selected from:
in the compounds used in the process of the invention, L is selected from the group consisting of a bond, C1-C6Alkyl radical, C2-C6Alkenyl radical, C1-C6Alkoxy radical, C1-C6Alkoxy radical C1-C6Alkyl and C1-C6A heteroalkyl group.
In one embodiment, L is a bond. In one embodiment, L is C1-C6An alkyl group. In one embodiment, L is C2-C6An alkenyl group. In one embodiment, L is C1-C6An alkoxy group. In one embodiment, L is C1-C6Alkoxy radical C1-C6An alkyl group. In one embodiment, L is C1-C6A heteroalkyl group.
In one embodiment, L is C of the formula1-C6Alkyl groups:
-(CH2)a-；
wherein a is selected from 1, 2, 3 and 4.
In one embodiment, a is 1 and L is-CH2-. In one embodiment, a is 2 and L is- (CH)2)2-. In one embodiment, a is 3 and L is- (CH)2)3-. In one embodiment, a is 4 and L is- (CH)2)4-。
In the compounds used in the process of the invention, R1Selected from H, OH, CN, tetrazole, CO2H and COR2。
In one embodiment, R1Is H. In one embodiment, R1Is OH. In one embodiment, R1Is CN. In one embodiment, R1Is tetrazole. In one embodiment, R1Is CO2H. In one embodiment, R1Is COR2。
In the compounds used in the process of the invention, R2Selected from H, Cl, NR3R4、O-C1-C6Alkyl and O-C1-C6A heteroalkyl group.
In one embodiment, R2Is H. In one embodimentIn the scheme, R2Is Cl. In one embodiment, R2Is NR3R4. In one embodiment, R2Is O-C1-C6An alkyl group. In one embodiment, R2Is O-C1-C6A heteroalkyl group.
In the compounds used in the process of the invention, R3And R4Each independently selected from H and C1-C6An alkyl group. In one embodiment, R3Is H. In one embodiment, R3Is C1-C6An alkyl group. In one embodiment, R3Is CH3. In one embodiment, R4Is H. In one embodiment, R4Is C1-C6An alkyl group. In one embodiment, R4Is CH3。
In the compounds used in the process of the invention, each R is5Independently selected from H, halogen, OH, NO2、CN、SH、NH2、CF3、OCF3、C1-C12Alkyl radical, C1-C12Alkoxy radical, C1-C12Haloalkyl, C2-C12Alkenyl radical, C2-C12Alkynyl, C2-C12Heteroalkyl, SR6、SO3H、SO2NR6R6、SO2R6、SONR6R6、SOR6、COR6、COOH、COOR6、CONR6R6、NR6COR6、NR6COOR6、NR6SO2R6、NR6CONR6R6、NR6R6And an acyl group;
or any two R on adjacent carbon atoms5When taken together with the carbon atom to which they are attached form a 5 or 6 membered ring moiety;
each R6Independently selected from H and C1-C12An alkyl group.
In one embodiment, each R is5Independently of each otherSelected from H, Cl, Br, F, OH, NO2、NH2、C1-C12Alkyl radical, C1-C12Alkoxy and NR6COR6。
In one embodiment, each R is5Independently selected from H, F, Cl, Br, I, CH3、CH2CH3、CH2NH2、OH、OCH3、SH、SCH3、CO2H、CONH2、CF3、OCF3、NO2、NH2CN and NHCOCH3。
In certain embodiments of the invention, the compound used in the method is such that X is S, X1Is O, X2Is O; two R when taken together form a double bond, R1Is CO2H, and Ar is a group of the formula:
this provides a compound of formula (4):
l, A therein1、A2、A3、A4And A5As defined above.
In the compounds of the formula (4) used in the process according to the invention, A1、A2、A3、A4And A5Each independently selected from N and CR5。
In one embodiment, A is1、A2、A3、A4And A5Each is CR5This provides a compound of formula (5).
Wherein L and R5As defined above.
In the formula 5In certain embodiments of (A), L is-CH2-. This provides a compound of formula (6):
wherein R is5As defined above.
Some examples of specific compounds of formula (1) for use in the process of the invention include the following or salts or N-oxides thereof:
the compounds of the present invention disclosed above are capable of inhibiting lysine biosynthesis in an organism in which the diaminopimelate biosynthetic pathway occurs by contacting the organism with an effective amount of the compound. Accordingly, the present invention also provides a method of inhibiting lysine biosynthesis in an organism in which the diaminopimelate biosynthetic pathway occurs, said method comprising contacting the organism with an effective amount of a compound of formula (1):
the organism is typically contacted with the compound of formula (1) by contacting the organism with a composition comprising the compound. In addition to the compounds, the compositions typically comprise a suitable solvent or carrier as detailed below for the herbicidal composition. The concentration of the compound of formula (1) in the composition may vary, but it is typically from 50 micromolar to 4000 micromolar. In one embodiment, the concentration is from 50 micromolar to 2000 micromolar. In one embodiment, the concentration is from 50 micromolar to 1000 micromolar. In one embodiment, the concentration is from 100 micromolar to 1000 micromolar. In one embodiment, the concentration is from 200 micromolar to 1000 micromolar. As will be appreciated by those skilled in the art, higher concentrations will work, but the higher the concentration, the more expensive the treatment becomes.
The organism may be any organism in which lysine biosynthesis takes place via the diaminopimelic acid pathway. In one embodiment, the organism is selected from the group consisting of plants and bacteria. In one embodiment, the organism is a plant. In another embodiment, the organism is a bacterium. In one embodiment, the organism is a gram-positive bacterium. In one embodiment, the organism is a gram-negative bacterium.
Without wishing to be bound by theory, it is believed that the compounds of the present invention inhibit lysine biosynthesis in organisms by inhibiting the diaminopimelic acid pathway. Thus, in some embodiments, the compounds inhibit lysine biosynthesis in an organism by inhibiting the diaminopimelate pathway. In some embodiments, the compound inhibits lysine biosynthesis in an organism by inhibiting DHDPS activity.
In inhibiting lysine biosynthesis, the compounds of the present invention are generally used in the form of compositions. In one embodiment, the composition is a herbicidal composition as discussed below.
Herbicidal composition
The herbicide composition comprising the active agent may be in the form of a liquid or solid composition, and thus the composition may be in the form of a concentrate, wettable powder, granules, or the like. Generally, these are intended to be mixed with other substances prior to application as herbicides. In these formulations, the active agent is typically present in 1 wt% to 90 wt% based on the total weight of the composition, with the remainder of the composition being made up of solid or liquid carriers and other additives as discussed below. In one embodiment, the active agent is present at 0.1 wt% to 90 wt% based on the total weight of the composition. In one embodiment, the active agent is present at 0.1 wt% to 50 wt% based on the total weight of the composition. In one embodiment, the active agent is present at 0.1 wt% to 10 wt% based on the total weight of the composition. In one embodiment, the active agent is present at 0.1 wt% to 5 wt% based on the total weight of the composition. In one embodiment, the active agent is present at 0.1 wt% to 1 wt% based on the total weight of the composition. In one embodiment, the active agent is present at 0.1 wt% to 0.5 wt% based on the total weight of the composition.
As will be appreciated by those skilled in the art, the concentration of active compound in the composition for contact with plants may vary widely depending on a number of factors. In one embodiment, the concentration is above 31.3 micromolar. In one embodiment, the concentration is above 62.5 micromolar. In one embodiment, the concentration is greater than 125 micromolar. In one embodiment, the concentration is greater than 250 micromolar. In one embodiment, the concentration is greater than 500 micromolar. In one embodiment, the concentration is greater than 1000 micromolar. In one embodiment, the concentration is from 15.6 micromolar to 500 micromolar. In one embodiment, the concentration is from 31.3 micromolar to 2000 micromolar. In one embodiment, the concentration is from 62.5 micromolar to 2000 micromolar. In one embodiment, the concentration is from 125 micromolar to 2000 micromolar. In one embodiment, the concentration is from 125 micromolar to 1000 micromolar. In one embodiment, the concentration is 250 micromolar to 1000 micromolar.
Suitable solid carriers for use in the herbicidal compositions include, but are not limited to: clays such as kaolinite, diatomaceous earth, synthetic hydrated silica and bentonite; talc and other inorganic substances such as calcium carbonate, activated carbon, powdered sulfur and powdered quartz; and inorganic fertilizers such as ammonium sulfate, ammonium nitrate, ammonium chloride, and the like.
Suitable liquid carriers may include: water; alcohols such as methanol, ethanol, 2-ethylhexanol, and n-octanol; halogenated hydrocarbons such as dichloroethane and trichloroethane; aromatic hydrocarbons such as toluene, xylene, and ethylbenzene; non-aromatic hydrocarbons such as hexane, cyclohexane, and the like; ketones such as acetone, methyl ethyl ketone and cyclohexanone; esters such as ethyl acetate and butyl acetate; nitriles such as acetonitrile, isobutyronitrile, and the like; ethers such as dioxane and diisopropyl ether; amides such as dimethylformamide and dimethylacetamide; or an organic sulfur compound such as dimethyl sulfoxide. In some embodiments, the liquid carrier is a mixture of one or more of these substances.
The composition may comprise one or more additional additives such as surfactants, crystallization inhibitors, viscosity modifying substances, suspending agents, dyes, antioxidants, foaming agents, light absorbers, mixing aids, antifoaming agents, complexing agents, neutralizing or pH adjusting substances and buffers, corrosion inhibitors, fragrances, wetting agents, absorption modifiers, plasticizers, lubricants, dispersants, thickeners and the like.
Surfactants useful in the herbicidal compositions of the present invention are well known in the art and include: salts of alkyl sulfates, such as diethanolammonium lauryl sulfate; salts of aryl sulfonates such as calcium dodecylbenzenesulfonate; alkylphenol-alkylene oxide addition products, such as nonylphenol ethoxylate; alcohol-alkylene oxide addition products, such as tridecyl alcohol ethoxylate; soaps, such as sodium stearate; salts of alkylnaphthalene sulfonates, such as sodium dibutylnaphthalene sulfonate; salts of dialkyl sulfosuccinates, such as sodium bis (2-ethylhexyl) sulfosuccinate; sorbitol esters, such as sorbitol oleate; quaternary amines, such as lauryl trimethyl ammonium chloride; polyethylene glycol esters of fatty acids, such as polyethylene glycol stearate; block copolymers of ethylene oxide and propylene oxide; and salts of monoalkyl phosphates and dialkyl phosphates.
Further additives which may be present in the herbicidal composition are those well known in the art. Herbicidal compositions are typically prepared by combining each of the desired ingredients into a formulation mixer and mixing to produce the final formulation.
Suitable herbicidal formulations comprising the compounds of formula (1) can be readily prepared by those skilled in the art of herbicidal formulation.
Use as herbicides
As mentioned above, the compounds of formula (1) are useful as herbicides. Accordingly, in one embodiment, the present invention provides a method for controlling undesired plant growth, which comprises contacting the plants with a herbicidally effective amount of a compound of formula (I) or a salt or N-oxide thereof.
Although in principle the compounds can be used to control the growth of any plant, they are generally used to control the growth of undesirable plants (e.g. weeds), particularly in an agricultural environment.
Some examples of plants that can be controlled using the methods of the present invention include: bindi (Bindii), Bindweed (Bindweed), cyperus brevifolia (Mullumbimby fruit), nettle (stinging nettle), panaspen (pampas grass), lantana (lantana), calendula (Capoweed), common sow (common sow tree), African wolfberry (African wolfberry tree), asparagus (asparagus fern), expectorant (asthmannia weed), black nightshade (black nightshade), blue morning glory (blue morning glory), broad-leaved marthria (brazilian crop), origanum (ox-eye daisy), sorrel (sorrel), lipstick (lippia liparis), purple nutgrass (purple grass), common sage (onion grass), bromeline (black plum), cockspur grass (tare), tarry (black plum), dandelion (black plum), black plum (black plum), black plum, Annual ryegrass (annual rye grass), Barley grass (Barley grass), buckwheat tendril (Black bindweed), bellflower (windbell kenmia), brome grass (kernel grass), spiny fruit (doublee), fleabane (fleabane), corydalis globosa (fungmitory), allium sativum (Indian hedge mulgard), lemongrass (Liverseed), musella foenum-graecum (muskeds), phalaris (Paradoxa grass), silvergrass (silvergrass), Sweet grass (Sweet summer grass), camelina (turnweed), Wild oat (Wild oat), Wild radish (Wild radish), windmills (windmills) and wirelines (wired).
The compounds of formula (1) may be applied to plants in any manner known in the art. However, the compounds are typically used in this method in the form of herbicidal compositions as discussed above. Applying a compound to a plant in this form typically involves applying a composition comprising the active agent to the plant as such, or by diluting the composition in a solvent (e.g., water) and then applying the diluted composition to the plant. Thus, application of a compound to plants typically involves contacting the plants with the compound in the form of a pure compound or a herbicidal composition. The compound may be applied by contact with any part of the plant, but this typically occurs through the roots, leaves or stems of the plant.
Application of the composition to the plant by contact may be by any method known in the art. Thus, for small scale applications, the composition comprising the compound may be manually applied or applied to the plant. For larger scale applications, the composition comprising the compound is typically applied by spraying (spraying), as is well understood by those skilled in the art. The amount applied (rate of application) will vary depending on the plants to be controlled, the amount applied (application rate), the maturity of the plants to be controlled and their degree of infestation of the land to be treated. In one embodiment, the application rate is typically from 0.1kg to 1000kg per hectare. In one embodiment, the application rate is from 0.1kg to 100kg per hectare. In one embodiment, the application rate is from 0.1kg to 50kg per hectare. In one embodiment, the application rate is from 10kg to 50kg per hectare. In one embodiment, the application rate is typically from 0.1kg to 50kg per hectare. In one embodiment, the application rate is from 0.1kg to 10kg per hectare. In one embodiment, the application rate is from 1.0kg to 0kg per hectare. In one embodiment, the application rate is from 1.0kg to 5kg per hectare.
The aqueous concentrate composition may be diluted in a suitable volume of water and applied to the unwanted plants to be controlled, for example by spraying. The compositions prepared by this method can be applied in amounts of, for example, about 0.1 to about 5 kilograms per hectare (kg/ha) (occasionally more). Typical amounts for controlling annual and perennial grasses and broadleaf grasses are from about 0.3 to about 3 kg/ha. The compositions of the present invention may be applied in any convenient volume of water, most typically from about 30 to about 2000 liters per hectare (l/ha) of water. Compositions useful in the methods of the present invention also include solutions that can be applied, for example, by spraying. In these solutions, the concentration of the active agent is selected according to the volume of spray solution used per unit area and the desired amount of active agent applied per unit area. For example, conventional spraying is carried out with spray liquors of 30 to 5000 litres (especially 50 to 600 litres) per hectare and the application rates of active agent are typically 0.125 to 1.5kg of active agent per hectare. Spray compositions may be prepared by diluting an aqueous liquid concentrate, preferably containing a surfactant adjuvant, or by tank mixing (tank mixing) the aqueous concentrate formed by this process with an adjuvant as described above.
Synthesis of Compounds of the invention
The compounds used in the methods of the present invention can be prepared using reaction schemes and synthetic schemes described below, using readily available starting materials using techniques available in the art. The preparation of specific compounds of some embodiments is described in detail in the following examples, but the skilled artisan will recognize that the chemical reactions described can be readily adapted to prepare many other reagents of the various embodiments. For example, the synthesis of non-exemplified compounds can be successfully carried out by modifications apparent to those skilled in the art: for example, by appropriately protecting interfering groups, by changing to other suitable reagents known in the art, or by making routine modifications to the reaction conditions. A list of suitable protecting Groups for Organic Synthesis can be found in T.W. Greene, Protective Groups in Organic Synthesis, 3 rd edition, John Wiley & Sons, 1991. Alternatively, other reactions disclosed herein or known in the art will be recognized as suitable for preparing other compounds of the various embodiments.
The invention will now be illustrated by way of examples; however, the examples should not be construed as limiting thereof. Additional compounds in addition to those described below may also be prepared using the methods and synthetic schemes as described herein or suitable variations or modifications thereof.
Most materials were purchased as reagent grade from Sigma-Aldrich. If they are not available from Sigma-Aldrich, they purchase from other commercial suppliers. The melting points obtained were uncorrected and recorded on a Reichert "Thermopan" microscope hot stage apparatus.
Nuclear Magnetic Resonance (NMR) spectra were obtained on a Bruker Avance-400 spectrometer1H at 400.13MHz and13c at 100.62 MHz) or Bruker Avance-500 spectrometer (1H at 500.03MHz and13c at 125.75 MHz). Proton chemical shifts are reported in ppm relative to residual chloroform (7.26ppm), dimethylsulfoxide (2.50ppm) or methanol (3.31ppm) internal standards. Each resonance is assigned according to the following convention; chemical shift (. delta.) (multiplicity, coupling constant: Hz, integration). Carbon chemical shifts are reported in parts per million (ppm) using internal standards of residual chloroform (77.16ppm), dimethyl sulfoxide (39.52ppm), or methanol (49.00 ppm). Chemical shifts are reported as delta values in parts per million (ppm). The following abbreviations were used in reporting the spectroscopic data: s, singlet; d, doublet; t, triplet; q, quartet; quin, quintuple peak; sext, sextuple peak; m, multiplet; app, appearance; and br, broad peak.
Electrospray ionization (ESI) Mass Spectrometry Using Bruker Daltonics (Germany) Esquire6000The ion trap mass spectrometer was operated at 140 ℃ in positive ion mode with a flow rate of 4 μ L/min, a mass range of 50m/z to 3000m/z and a scan rate of 5500 m/z/sec. Methanol was used, together with 0.1% formic acid as mobile phase.
Thin Layer Chromatography (TLC) was used to monitor reactions and chromatographic fractions on a Merck Kieselgel 60F254 aluminum back plate. Silica gel 60F254 was used as the stationary phase for flash chromatography. Unless otherwise stated, a gradient elution was performed using analytical grade ethyl acetate (EtOAc) and hexanes.
Analytical reverse phase High Performance Liquid Chromatography (HPLC) is performed in a cell Jupiter C18The columns (250 mm. times.4.60 mm, 10 μm) were performed on a Shimadzu research HPLC system using the following buffer binary system: solvent A: 0.1% trifluoroacetic acid; solvent B: and (3) acetonitrile. Gradient elution was performed using a gradient of 90% solvent a to 90% solvent B, monitored at 254nm over 20 minutes at a flow rate of 1 mL/min. Unless otherwise stated, semi-preparative reverse phase HPLC is carried out using a column packed with Jupiter C18The previously described system of columns (250 mm. times.10.0 mm, 10 μm) was carried out in 60 minutes at a flow rate of 2 mL/min using the same binary buffer system described for RP-HPLC.
All glassware for reactions requiring anhydrous conditions was dried (120 ℃) and subsequently cooled under nitrogen prior to use.
A general scheme for forming the compounds of the present invention is shown in scheme 1 below, which may be directed to Ar, X in the final product1、X2L and R1The variables selected to modify.
Generally, a suitably functionalized Ar-aldehyde (a) is reacted with a suitably functionalized heterocyclic group (e.g. 2, 4-dioxothiazolidine (when X ═ S, X) in the presence of traces of piperidine and acetic acid1＝O、X2O), 4-oxo-2-thioxothiazolidine (when X is S, X)1＝O、X2When X ═ S), hydantoin (when X ═ NH, X)1＝O、X2O ═ O) and thiohydantoin (when X ═ NH, X)1＝O、X2When ═ S)) (B) are reacted under reflux to form condensation product C. In the reaction, R on (B)1The groups are typically protected as esters of the free acid. As the skilled person will understand, X, X1And X2Other combinations of (a) can be performed using suitable starting materials. After condensation, the ester groups on (C) can be removed under acidic conditions to form free radicals, if desiredAnd (4) separating the substances.
Reagent B used in scheme 1 is typically generated as shown in scheme 2. Accordingly, a suitable heterocyclic amine (B1) is reacted with a suitable functionalizing reagent (B2) containing a suitable leaving group (in this case Br) under mild basic conditions to yield reagent B as used in scheme 1.
Almost all compounds of the present invention can be produced using the procedures described in the above reaction schemes with minor modifications within the ability of the organic synthesis chemist.
Synthesis of ethyl 2- (2, 4-dioxothiazolidin-3-yl) acetate (starting Material A)
To a stirred suspension of 2, 4-thiazolidinedione (0.200g, 1.71mmol) and potassium carbonate (0.473, 3.42mmol) in anhydrous acetonitrile (30mL) under nitrogen was added ethyl bromoacetate (0.208mL, 1.88mmol) dropwise. After stirring at room temperature for 18 h, the reaction was concentrated in vacuo and the residue was partitioned between ethyl acetate (20mL) and water (20mL), and the aqueous phase was extracted with ethyl acetate (3 × 20 mL). The organic phase was dried (MgSO)4) And concentrated. The crude product was subjected to column chromatography (silica; 20: 80 ethyl acetate/hexane elution) to give starting material A (0.279g, 80%) as a pale yellow oil.
δH(400MHz，CDCl3)4.35(s，2H，CH2)，4.23(q，J 16.0，8.0，2H，CH2)，4.04(s，2H，CH2)，1.29(t，J 8.0，3H，CH3).δC(100MHz，CDCl3)171.1，170.7，166.2，62.1，42.1，33.9，14.0.
Synthesis of 2- (2, 4-dioxothiazolidin-3-yl) acetic acid (starting Material C)
A mixture of starting material A (0.250g, 1.23mmol) in glacial acetic acid (6mL) and concentrated hydrochloric acid (3mL) was refluxed for 1 hour. The reaction was concentrated in vacuo and the residue was partitioned between water (20mL) and ethyl acetate (25 mL). The aqueous phase was washed with ethyl acetate (3X 25mL) and dried (MgSO)4) And concentrated in vacuo to give an oil that solidified in vacuo (0.183g, 85%).
δH(400MHz，DMSO)4.33(s，2H，CH2)，4.21(s，2H，CH2).
Example 1- (Z) -2- (5- (4-Fluorobenzylidene) -2, 4-dioxothiazolidin-3-yl) acetic acid
Step Synthesis of ethyl 1- (Z) -2- (5- (4-fluorobenzylidene) -2, 4-dioxothiazolidin-3-yl) acetate
To a solution of 4-fluorobenzaldehyde (0.211mL, 1.97mmol) and (starting material A) (0.400g, 1.97mmol) in toluene (8mL) was added six drops of piperidine and four drops of acetic acid. The reaction was heated at reflux for 18 hours, after which the yellow precipitate formed was allowed to cool. The precipitate was collected by vacuum filtration and washed with a small amount of toluene to give the desired compound (0.323g, 53%).
δH(400MHz，CDCl3)7.91(s，1H，CH)，7.53(dd，J 8.0，4.0，2H，ArH)，7.19(t，J 8.0，2H，ArH)，4.48(s，2H，CH2)，4.45(q，J 16.0，8.0，2H，CH2)，1，30(t，J 8.0，3H，CH3).δC(100MHz，CDCl3)167.2，166.2，165.5，165.1，162.5，133.3，132.4，132.3，129.42，129.39，120.8，120.7，116.7，116.5，62.2，42.2，14.1.
Step Synthesis of 2- (Z) -2- (5- (4-Fluorobenzylidene) -2, 4-dioxothiazolidin-3-yl) acetic acid
A mixture of ethyl (Z) -2- (5- (4-fluorobenzylidene) -2, 4-dioxothiazolidin-3-yl) acetate (0.270g, 0.873mmol), glacial acetic acid (12rnL) and concentrated hydrochloric acid (5mL) was refluxed for 2 hours. The reaction was concentrated in vacuo, and the product was washed with water and dried to give the desired compound (0.224g, 91%).
δH(400MHz，DMSO)13.42(br s，1H，COOH)，8.01(s，1H，CH)，7.73(dd，J 16.0，8.0，2H，ArH)，7.40(t，J 8.0，2H，ArH)，4.37(s，2H，CH2).δC(400MHz，DMSO)168.4，167.3，165.5，164.8，162.4，133.32，133.28，133.2，130.0，129.9，120.90，120.87，117.2，117.0，42.8.
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